Online Exclusive Article

PHARMACEUTICAL ENGINEERING
The Official Magazine of ISPE Risk Management in Sterile Facilities
March/April 2010, Vol. 30 No. 2

This article
presents Design, Validation, and Control of
the design,
validation, and Sterile Manufacturing Facilities: A
control of sterile
manufacturing Brief Overview from the Perspective
facilities;
discusses the
implementation
of Risk Management and Existing
of risk
management,
Regulations
and provides
an overview
of existing by Ana Quinto and Jos C. Menezes
regulations.

Introduction

O
processes less predictable, naturally having
ne of the most critical operations in higher risk, and being more difficult to control
pharmaceutical manufacturing is the and manage:
processing of sterile products. The pro-
duction of sterile products, specifically Aseptic manufacturing processes are unique
the ones that cannot be terminally sterilized, since the severity of harm is always going to be
involve complex and demanding processes to high and detection of loss of sterility is always
prevent the products contamination and require going to be low.1,2
a great amount of resources.
The inherent risk of microbiological con- To enter this business successfully, it is impor-
tamination associated with aseptic operations tant to have a deep knowledge of what underlies
is critical because it has a direct relation with this type of manufacturing, such as the strict and
human health. The difficulty in detecting extended regulations applicable and the cost of
contamination makes the outcome of these what is necessary to start and maintain these
Figure 1. Cause and
effect diagram with
microbiological
contamination
parameters from an
aseptic process.
Copyright ISPE 2010

www.ISPE.org/PE March/april 2010 PHARMACEUTICAL ENGINEERING Online Exclusive 1

 Risk Management in Sterile Facilities
types of processes. Adequate facilities, equipment, materials,
procedures, operators, and a strong and robust sterility as-
surance policy are examples of these requirements. The EU
GMP Annex 1 reinforces this idea referring that:
Sole reliance for sterility or other quality aspects must not be
placed on any terminal process or finished product test.1,2
Sterile products can be processed by aseptic manufacturing
or can be terminally sterilized. These two types of processes
have different characteristics and involve specific conditions.
The regulatory guidelines describe specific conditions expected
for each one regarding every step of the process.2,3
Terminally sterilized products involve manufacturing
processes, where microbiological contamination can happen
within highly controlled conditions. Sterility is obtained
through a final sterilization step, where the product is already
in its final container.2,3
Aseptic manufacturing is more demanding as there is no
final sterilization step.
Freeze-drying is a specific type of production process that
involves products that are unstable as solutions. In these
situations, the process involves the filtering of the solutions
before a filling step, adding the freeze-dry step that happens
with the containers opened. The final closure of the contain-
ers only occurs much later in the process, which represents
a great contamination risk.4
Sterility of products, particularly the ones produced by
aseptic processing, is obtained guaranteeing the conformity
of the processes different factors - Figure 1. To obtain sterile
products, it is essential that all the processing is done in a
way that minimizes the risk of contamination hazards.1,2
A contamination event in any of the referred factors in the
previous diagram is considered critical for the sterility of the
process. The concept of Quality by Design is discussed in the
FDAs Guidance for Industry: PAT:
Quality cannot be tested into products; it should be built-in
or should be by design.5
The changes in the industry in the last 10 years helped define
a strong and growing set of regulations and guidance docu-
ments related to this subject. These regulatory documents
define requirements supported by standards and industry
guidance published by groups, including ISPE and PDA.
Risk Management
Although for many years, the concept of risk management
also has been applied in the pharmaceutical industry in
an informal manner, formal applications are more recent
and still considered limited. The FDAs initiative in 2002,
Pharmaceutical CGMPs for the 21st Century: A Risk-Based
Approach was extremely important in promoting risk man-
agement since it presented the first structured approach in
this area.1 The main purpose of the use of risk management
is to support decisions using rational methodology although
it is important to underline that compliance with regulatory
2 PHARMACEUTICAL ENGINEERING Online Exclusive March/April 2010
aspects continues to be a requirement.1,6
Quality risk management was defined in the guideline
ICH Q9 as a systematic process for the assessment, control,
communication, and review of risks to the quality of the drug
(medicinal) product throughout its lifecycle.6,7
An adequate use of quality risk management tools provides
better and more informed decisions, enabling, for example,
regulators to assert a companys capability to deal with risk
problems and positively affect the thoroughness of direct
regulatory supervision. The main purpose of the use of risk
management is to support decisions using rational method-
ology although it is important to underline that compliance
with regulatory aspects continues to be a requirement.1,6
Facilities
Most sterile manufacturing processes are performed in clean-
rooms. A cleanroom can be defined as a room in which the
concentration of airborne particles and other environmental
conditions, such as temperature, humidity, and pressure, are
controlled.9
This type of environment is a requirement for the manu-
facturing of sterile products in order to minimize the risk of
microbiological, particle, and pyrogen contamination.2,3 The
air handling and the type of surface materials are examples
of important issues to be dealt with in the construction of a
cleanroom. The necessary space must be available in clean-
rooms so all the operations and procedures can be performed
properly. Equipment and other items introduced in cleanrooms
should be considered since they can influence the cleanrooms
performance.4
The air that enters the cleanroom has to be filtered by
an adequate High Efficiency Particulate Air (HEPA) filter,
in order to prevent contaminations from the outside. The
number of air changes has to be adequate to dilute contami-
nation generated from the process; equipment and personnel
and airflow patterns inside the clean areas must prevent the
contamination of the critical areas, where sterile products
and other important items are manipulated. The pressure
differentials inside the aseptic facility must prevent airflows
from less clean areas to cleaner areas. When dealing with
potent parenteral products, the pressurization scheme also
must address containment issues.4
Cleanrooms can be divided in four types, according to the
airflow pattern:
Conventional
Unidirectional Flow
Mixed Flow
Isolators, RABS, or Microenvironments11
Isolators, RABS (open or closed), or microenvironments are
Copyright ISPE 2010
the ones that offer better processing conditions since access
to the critical area is very limited. These critical areas, where
sterile materials are exposed, are completely segregated and
protected by unidirectional flow.11
Mixed flow is considered the basic design concept for a
cleanroom with a unidirectional flow inside a conventional
www.ISPE.org/PE
 Risk Management in Sterile Facilities
room with turbulent flow. In this case, the critical areas are
not as protected as the previous situations.11
Unidirectional flow cleanrooms are completely covered
by unidirectional flow. In this case, the critical areas are not
segregated from other areas.11
Conventional cleanrooms are the ones that present less
protection for the product, as the critical areas also are not
segregated from other areas and the flow inside the room is
turbulent.11
In situations where the product is less protected, very well
defined procedures are required to prevent contamination.11

Cleanroom Classification
Regulatory documents and norms define tests and specifi-
cations to demonstrate the compliance of the cleanroom to
certain cleanliness classes. EU and FDA GMPs refer to ISO
Standards in what concerns detailed methods for classifica-
Figure 2. Graphical illustration of the airborne particulate
tion of cleanrooms.2,3,4 cleanliness classes.9
Parameters that must be evaluated when testing a clean-
room involve: where, Cn is the maximum permitted concentration (particles
per m3) of particles, equal to or larger than the considered
leak tests to the HEPA filters particle size, N is the ISO classification number, D is the
number of particles considered particle size in micrometers, 0.1 is a constant with
air change rates a dimension of micrometers.9
recovery times of the cleanroom after a contamination Figure 2 has a graphical representation of airborne par-
event ticulate classes obtained using Equation 1.9
airflow patterns The last version of the EU GMPs still presents some dif-
pressure differentials ferences in these specifications compared to the ISO and the
FDA. Table A has the concentration limits of airborne particles
Nevertheless, the number of total particles is one of the main for each grade of cleanliness, according to the latest version
issues when classifying a cleanroom. ISO and FDA particle of Annex 1 EU GMP,1,2 ISO 14644-1,9 and FDA guidance,3
limits for classification purposes are determined from the where it is possible to observe the differences and similarities
following Equation 1.9 between their specifications.
Regarding these limits, the major controversy happened
0.1 2.08
with the 5 m limits, as the EU limit in the 2003 version of
Cn = 10 ______
N

D the guideline, was 1/m3 a number not achievable given the

measuring limitations.7,12 In the 2005 GMP Annex 1 propos-

Guideline Grade At Rest In Operation

0.5 m 5.0 m 0.5 m 5.0 m
EU A / B** 3 520 20 / 29 3 520 20
ISO* 5 3 520 29 3 520 29
FDA 100 --- --- 3 520 ---
EU --- --- --- --- ---
ISO* 6 35 200 293 35 200 293
FDA 1000 --- --- 35 200 ---
EU B*** --- --- 352 000 2 900
ISO* 7 352 000 2 930 352 000 2 930
FDA 10 000 --- --- 352 000 ---
EU C 352 000 2 900 3 520 000 29 000
ISO* 8 3 520 000 29 300 3 520 000 29 300
Copyright ISPE 2010

FDA 100 000 --- --- 3 520 000 ---

EU D 3 520 000 29 000 Not defined Not defined
ISO* 9 35 200 000 293 000 35 200 000 293 000
FDA --- --- --- --- ---
*ISO designation should include the classification number and the occupancy state to which the classification applies as-built, at rest, and operational, as
there is no other discrimination in the limits.8 **Refers to grade B at rest. ***Refers to grade B in operation.
Table A. EU, ISO, and FDA maximum permitted number of particles per m3 for each grade.2,3,9

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 Risk Management in Sterile Facilities
als of amendment, a limit of 20/m3 was suggested for reasons particles that could be counted if the particle concentration
related to false counts associated to electronic noise.13 This were at the class limit.15
limit of 20/m3 for the 5 m particles was officially adopted Nevertheless, the ISO document is currently going through
in the latest version of 2008. The other EU limits also were its periodic review with some sections being modified. The
slightly different from those of the ISO.1,2,3,9 number of sampling points proposed in ISO 14644-1 for clas-
Although smaller, the differences in the limits presented sification purposes was adopted both by EU and FDA, which
by the current EU guidelines and the ISO/FDA still exist, but is the number corresponding to the areas square root.1,2,15
there is a strong international pressure to end the differences, Microbiological limits also are a critical issue when
and obtain a global harmonization.7,14 The major differences evaluating pharmaceutical production cleanrooms.1,2,3 The
between the current EU particle limits, the ISO, and the FDA manufacturers are expected to have standards based on
are: their processes historic values. The characterization of the
microorganisms also is an expectation.
FDA only considers 0.5 m particle limits Table B presents the microbiological limits in Colonies
EU 5 m limits (20/m3) are more rigorous regarding the Forming Units (CFU) for each grade of cleanliness, accord-
ISO limits (29/m3) ing to the latest version of Annex 1 EU GMP (2008) and the
FDA only considers limits for operation conditions, EU has FDA.1,2,3 Microbiological limits in both guidelines are similar,
limits for at rest and in operation, and ISO has only the following are the major differences between the EU and
one type of limits, meaning that ISO designation should the FDA guideline:
include the classification number and the occupancy state
to which the classification applies as-built, at rest, EU allows average numbers, which is a controversial topic
and operational between the two agencies, since it is much less restrictive
FDA/ISO consider an intermediate classification level than the individual limits presented by the FDA. This is-
between 100/ISO 5 (EU grade A) and 10 000/ISO 7 (EU sue is particularly relevant in Grade A/Class100.
grade B), which is Class 1000/ISO 6. This level can be used, FDA has limits for Class 1000.
for example, for areas surrounding Class 100. FDA does not present limits for contact plates or gloves al-
FDA does not consider the cleanliness level corresponding though referring that operators gloves and gowns involved
to EU grade D.1,2,3,9 in aseptic operations, should be contamination-free.
FDA considers settling plates as an optional type of moni-
Another controversial issue concerned the minimum sample toring.1,2,3
volume of 1 m3 for cleanrooms classification purposes referred
to in the EU guideline, which is a much higher value compared Regarding microbiological limits, there were no changes in-
to the ISO values (view Equation 2).1,2,15 troduced by the 2008 version of the EU Annex 1.1,2,12,13
The ISO documents were not referred as they do not
20 present any microbiological limits, only referring tools and
Vs = ______ 1000
Cn,m methodologies for biocontamination control.16,17

The regulatory guidelines define expected minimum classi-
where, Vs is the minimum single sample volume per loca- fication conditions for each step of sterile production processes,
tion, expressed in liters, Cn,m is the Class limit (number of which must be considered when designing a manufacturing
particles per cubic meter) for the largest considered particle process.
size specified for relevant class, 20 is the defined number of
Ventilation and Cleanroom Design
Guideline Grade** Air Sample ToSettle
achieve the required
Plates classification
Contact Plates in a cleanroom,
Glove Point it is
(CFU/m )
3
diameter 90 mm diameter 55 mm 5 fingers
(CFU/4 hours) (CFU/plate) (CFU/glove)
EU A <1 <1 <1 <1
FDA 100 1* 1* --- ---
EU --- --- --- --- ---
FDA 1000 7 3 --- ---
EU B** 10 5 5 5
FDA 10 000 10 5 --- ---
EU C 100 50 25 ---
Copyright ISPE 2010

FDA 100 000 100 50 --- ---

EU D 200 100 50 ---
FDA --- --- --- --- ---
*No microbial contamination is expected on samples from Class 100.
**The association between EU and FDA grades was performed assuming in operation values (view Table A).
Table B. EU and FDA recommended limits for microbial contamination.2,3

4 PHARMACEUTICAL ENGINEERING Online Exclusive March/April 2010 www.ISPE.org/PE


Figure 3. Shell-like contamination control concept.19
necessary to install the capacity for the necessary airflow
rate, considering each cleanrooms particle generation rate.
Entries and exits of air should be installed according to the
room characteristics, equipments layout, and the production
processes that will be performed.18
The airflow between different areas must prevent con-
tamination of cleaner areas. Figure 3 presents a shell-like
contamination control concept with a progressive protection
of the process core at which the most critical operations oc-
cur.19 Like referred earlier, when dealing with potent products,
the pressurization scheme is different and more complex, as
containment issues also must be addressed.
Material transport inside the clean zone is accomplished
through sterilization/depyrogenation/sanitization processes.
Personnel movement regarding the clean zone is usually
done using gowning zones with several steps, and separated
entrance and exit areas.19 The final steps of the gowning
zones must be of the same grade as the areas where people
are going to enter.1,2,3
In the most critical areas, where sterile items are exposed,
Grade A/Class 100 environment is a requirement. Unidirec-
tional flow is a condition referred to in the guidelines for
Grade A/Class 100.1,2,3 The aim of this unidirectional flow is
to prevent any contamination entering the clean space and
also to remove any contamination from the critical areas as
fast as possible.
The regulatory documents refer minimum pressure differ-
entials that must be met between areas with different classifi-
cation. Pressure differentials of 10 to 15 Pa can be considered
guidance values for both EU and FDA guidelines.1,2,3
The number of change rates and locations of entries and
exits of air are very critical issues that deeply influence the
performance of a clean process, which must be established
for each particular situation. The location of Grade A/Class
100 areas also must be considered. Regarding these issues,
Copyright ISPE 2010
the FDA only refers 20 air changes per hour as an acceptable
number for Class 100000 number being challenged by many
people in the industry, and higher change rates for superior
cleanliness Classes.3 The EU GMPs refer that at rest limits
should be achieved after a short recovery period of 15 to 20
minutes after operations occur.1,2
www.ISPE.org/PE
Risk Management in Sterile Facilities
Practical study of the airflow patterns with smoke and
determining recovery times using real simulation models can
be extremely useful when establishing the necessary condi-
tions of a clean area.18,21 Another option to determine these
cleanroom parameters is the use of computer simulation mod-
els, such as, airflow design methods that allow incorporating
the referred performance variables. The Dilution Model and
Computational Fluid Dynamics are examples of methods for
airflow design.20,22
The accurate determination of air supply also concerns
energy savings, as these areas are highly energy consum-
ing.23 In this context, ISO 14644-4 also refers the possibility
of reducing air supply on cleanrooms during non-operating
periods to reduce costs.19
Equipment
Equipment and all other items used in aseptic processing areas
have specific requirements to allow the compliance with the
necessary environmental classifications. Characteristics like
the type of materials that must be non-shedding and shape
of surfaces are crucial in order to comply with the necessary
conditions and to enable an adequate cleaning and disinfec-
tion. Appropriate equipment design can prevent turbulence
and stagnant air in the critical areas. Equipments layout
inside the aseptic processing areas also should be addressed.
The components that are going to be in direct contact with
the sterile product must be sterilizable.1,2,3
The equipment and manufacturing process must be de-
signed and operated in a way that prevents contamination.1,2,3
All the required characteristics deemed important when
designing a process or choosing equipment should be clearly
defined and documented regarding the processes needs and
regulatory requirements.
Utilities
All utilities supplied to sterile processes, like any other item
entering the processes, must guarantee that the required
conditions for each step of the process are not disturbed.1,2,3
Validation and monitoring of utilities is a critical issue.24
Examples of such utilities, besides the air introduced into the
cleanrooms, are water, steam, and gases (e.g., compressed air
or nitrogen). The quality required is increasingly demanding
when closer to the critical areas.
Water to be used in the critical steps of sterile manu-
facturing must comply with the requirements of Water for
Injection (WFI). Examples of such use are injectable product
preparations, preparation of injections, final rinse after clean-
ing equipment and components that come into contact with
injectable products, and final rinse of a washing process in
which no subsequent thermal or chemical depyrogenization
process is applied.26 The European Pharmacopoeia is more
demanding in what concerns production of WFI, as it only
considers acceptable distillation as a production process.27
The USP allows other type of production processes.28 The most
critical issues in the production and distribution of water for
pharmaceutical use, and particularly WFI, are related to mi-
crobiological and endotoxin contamination control, involving
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 Risk Management in Sterile Facilities
special requirements.25
Specifications of steam to contact a sterile product or com-
ponent should comply with the specifications for WFI when
condensed.26
The production of medicinal gases is usually a specialized
industrial process, which is not normally undertaken by
pharmaceutical companies.29,30
Validation of Sterile Processes
Process validation is defined in the Annex 15 of the EU Guide
to GMP as the documented evidence that a process, operated
within established parameters, can perform effectively and
reproducibly to produce a medicinal product meeting its pre-
determined specifications and quality attributes.31
Classic process validation concepts are being replaced
by a new validation strategy based on process and product
understanding throughout the entire products lifecycle, par-
ticularly after the FDA initiatives Pharmaceutical CGMPs
for the 21st Century: A Risk-Based Approach.32 Science-based
methodologies and modern technology, such as, PAT5 and
risk-based approaches,7 were strongly encouraged for process
validation, monitoring, and control. The release of products
without having to perform all the tests in the specifications,
based on the information collected from the manufacturing
process and on the compliance with other specific GMP com-
pliance related to parametric release,33 is only considered in
this context of sterile manufacturing for terminally sterilized
products, based on the compliance of the critical parameters
of sterilization.34
A multi-disciplinary team is essential when dealing with
validations of sterile processes. Engineering, production,
quality assurance, microbiology, validation, and product
development are examples of people who must be involved.
The planned validation steps should be formally concluded
before moving to the next step. Process validation must be
completed prior to the distribution and sale of the medicinal
product. Revalidation of the processes should be addressed
regarding each specific situation to guarantee that the pro-
cesses remain valid, considering the situation when process
modifications occur.31
Qualification
Qualification can be defined as a documented scientific
process, used by pharmaceutical manufacturers, to assure
the reliability and capability of equipment and/or processes
before approval for use in manufacturing. Examples of pro-
cessing equipment that should be qualified in this aseptic
processing context are sterilizers, washing equipment, filters,
fillers, closure placement equipment, sealing machinery, and
freeze-dryers.35
A risk-based approach should be considered throughout
all the qualification phases. The establishment of monitor-
ing programs also should be based on a risk-based approach
considering the data collected during the qualification. Smoke
studies, showing air flow distribution and particle data, are
valuable information to be considered, particularly when
determining the monitoring locations.3 The determination
6 PHARMACEUTICAL ENGINEERING Online Exclusive March/April 2010
of the time for monitoring should consider critical activity
timings and production contingencies.
Process Simulation Testing
The most challenging issue, when validating a sterile produc-
tion process, is the microbiological contamination control, as
this type of process is expected to guarantee zero contamina-
tion. Other quality parameters specific of this type of product,
such as total particle counts and pyrogens, also are important,
but their limits and the experience indicates that they are
much easier to comply with.
To demonstrate that a certain manufacturing process
can consistently produce a sterile product, it is necessary to
assess the production system throughout the simulating of
the manufacturing process using a nutrient medium. This
simulation process using a nutrient medium, instead of the
real product, is usually called media fill.35
The latest version of the EU GMP Annex 11,2 and the
FDA guideline regarding sterile production,3 present similar
guidance regarding the number of containers that should be
filled to simulate the processes, frequency of tests, and also
acceptance criteria - Table C. Both documents refer that the
contamination goal of media fills should be zero.
Initial validation involves three consecutive satisfactory
simulation tests per shift. Media fills should be repeated at
defined intervals (e.g., twice a year) and when significant
process modifications occur.1,2,3
When designing the simulation test, it is necessary to select
worst-case scenarios (e.g., maximum number of operators, po-
tential interventions in critical areas, and time of production
process).35 Risk assessment tools are useful to help determine
the validation process. All operations must be included, such
as, compounding or filling. When simulating a powder filling
production line, the simulation test must be performed allow-
ing the same type of evaluation, for example, using a powder
placebo and adding a step in the filling process where media
is inserted into the container.35
The protocol should include all the information describ-
ing the process simulation test and supporting the choices
made. Identification of the process and operators, number of
containers being filled, type of containers, speed of the filling
line, interventions, type and amount of media and placebo,
Production Minimum Results and Required Actions
Batch Size Containers
(Containers) Tested Per Run
< 5000 Size of production No contaminated units should be
batch detected.
One (1) contaminated unit
investigation and revalidation.
5000 5000 10 000 One (1) contaminated unit
investigation and consideration of a
repeated media fill.
Copyright ISPE 2010
Two (2) contaminated units
investigation and revalidation.
10 000 One (1) contaminated unit
investigation.
Two (2) contaminated units
investigation and revalidation.
Table C. Media fill number of containers and acceptance criteria.2,3
www.ISPE.org/PE

duration of filling, environmental monitoring, acceptance
criteria, incubation conditions, rejected units, and results
are examples of information that should be included in such
protocols.35
This test is an important tool to have an idea of the aseptic
process capability, including environment, equipment, proce-
dures, and personnel. It does not assure the sterility of all
products produced in the tested manufacturing process, but in
combination with proper control of the processes (e.g., routine
monitoring program, validation, and personnel qualification),
it is possible to have an acceptable level of sterility assurance
regarding the aseptic processes.35
Maintaining Compliance
Control of the implemented processes is critical to assure the
maintenance of the installed conditions. A Routine Monitor-
ing Program (RMP) of an aseptic process intends to evaluate
the aseptic processes performance; therefore, the parameters
tested and the information acquired in the performance quali-
fication phase should be considered in the risk assessment
when developing this program.24,35
Regarding the referred parameters being monitored, a
RMP should include:
monitoring spots
frequency of monitoring
duration of sampling
when to sample
monitoring methods
alert and action levels
actions to be taken when limits are exceeded3,35
It is important to develop a proper management system to
help deal with the collected data and ease the evaluation
process.35
Conclusions
The design, validation, and control of sterile manufactur-
ing facilities were reviewed considering the most relevant
regulatory guidelines, applicable ISO documents, and other
significant references. A reference regarding implementation
of risk management in the context of sterile manufacturing
was presented. The most important issues that must be
considered in this type of facilities were discussed, including
the types of cleanrooms, cleanroom classification, ventilation
and design, equipment, utilities, validation, qualification, and
maintenance. Although it is clear that the trend throughout
the world is to harmonize regulations, the main differences
concerning the EU and the FDA GMPs were highlighted.
References
Copyright ISPE 2010
1. PDA Technical Report No. 44, Quality Risk Management
for Aseptic Processes, PDA Journal of Pharmaceutical
Science and Technology, 62(S-1) Supplement, 2008.
2. EU Guidelines to Good Manufacturing Practice, Medici-
nal Products for Human and Veterinary Use, Revision
to Annex 1: Manufacture of Sterile Medicinal Products,
www.ISPE.org/PE
Risk Management in Sterile Facilities
European Commission, Brussels, February 2008. http://
ec.europa.eu/enterprise/pharmaceuticals/eudralex/vol-4/
pdfs-en/2008_02_12_gmp_annex1.pdf.
3. Guidance for Industry, Sterile Drug Products Produced
by Aseptic Processing Current Good Manufacturing
Practice, FDA, Rockville, September 2004, http://www.
fda.gov/downloads/Drugs/GuidanceComplianceRegula-
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4. Mller, A., Chapter 2 International Standards for the
Design of Cleanrooms, in Whyte W., Cleanroom Design,
2nd Edition, John Wiley and Sons Ltd, England, 1999, pp.
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5. Guidance for Industry: PAT A Framework for Innovative
Pharmaceutical Development, Manufacturing, and Quality
Assurance, FDA, Rockville, September 2004. http://www.
fda.gov/CDER/guidance/6419fnl.pdf.
6. EU Guidelines to Good Manufacturing Practice, Medicinal
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ity Risk Management, European Commission, Brussels,
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7. ICH Q9 - Risk Management, Technical Report, Interna-
tional Conference on Harmonization, November 2005.
http://www.ich.org/LOB/media/MEDIA1957.pdf, 2008.
8. Farquharson, G., Using New Cleanroom Standards with
Sterile GMPs, Pharmaceutical Engineering, November/
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ronments Part 1: Classification of Air Cleanliness, ISO
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10. ISPE Baseline Pharmaceutical Engineering Guide, Vol-
ume 3 Sterile Manufacturing Facilities, International
Society for Pharmaceutical Engineering (ISPE), First
Edition, January 1999, www.ispe.org.
11. Whyte, W., Chapter 1 An Introduction to the Design of
Clean and Containment Areas, in Whyte, W., Cleanroom
Design, 2nd Edition, John Wiley and Sons Ltd, England,
1999, pp. 9-20.
12. EC Guide to Good Manufacturing Practice Revision to
Annex 1: Manufacture of Sterile Medicinal Products, Eu-
ropean Commission, Brussels, May 2003, http://ec.europa.
eu/enterprise/pharmaceuticals/eudralex/vol-4/pdfs-en/
revan1vol4_3.pdf.
13. GMP Annex 1: Proposals for amendment to the environ-
mental classification table for particles and associated text,
amendment to section 42 concerning acceptance criteria for
media simulations, amendment to section 52 concerning
bio-burden monitoring, and additional guidance in section
88 on the sealing of vials, (EMEA/INS/GMP/318222/2005/
Correction), EMEA Inspections, September 2005, http://
ec.europa.eu/enterprise/pharmaceuticals/pharmacos/docs/
doc2005/11_05/annex_1_amended_09-2005.pdf.
14. Valle, M., USA/EC/ISO Regulatory Considerations for
Designing Aseptic Processing Facilities, Pharmaceutical
Engineering, July/August 2004, Vol. 24, No. 4, pp. 36-46,
www.ispe.org.
March/april 2010 PHARMACEUTICAL ENGINEERING Online Exclusive 7
 Risk Management in Sterile Facilities
15. ISO 14644-2, Cleanroom and Associated Controlled
Environments Part 2: Specifications for Testing and
Monitoring to Prove Continued Compliance with ISO
14644-1, ISO 2000.
16. ISO 14698-1, Cleanrooms and Associated Controlled En-
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Acknowledgments
The authors would like to acknowledgment the National Inno-
vation Agency (ADI) in Portugal for financial support (SFRH/
BDE/15520/2004) and Laboratrios Atral, SA for providing
the best possible conditions to carry out this work, including
financing and permission for its publication.
About the Authors
Ana M. Quinto is the Sterility Assurance
Group Lead at Laboratrios Atral, SA. She
is a PhD student at the Technical University
of Lisbon (Portugal). The focus of her PhD
work is quality assurance of sterile products.
She received her university Pharmacist de-
gree in 2001 from the University of Lisbon,
Portugal, after which she started her career
in the pharmaceutical industry as production manager of
an aseptic facility. She continued her work in the Quality
Assurance Department, where she is currently Quality As-
surance Manager. Quinto has been engaged in environmental
monitoring of cleanrooms, validation, risk assessment, and
development of new production facilities. She is a member of
the Portuguese Society of Pharmacists. She can be contacted
by email: amquinto@atralcipan.pt.
Copyright ISPE 2010
Institute for Biotechnology and Bioengineering, Centre
for Biological and Chemical Engineering, IST, Technical Uni-
versity of Lisbon, Laboratrios ATRAL SA, Rua da Estao,
42, Vala do Carregado, P-2600-726 Castanheira do Ribatejo,
Portugal.
www.ISPE.org/PE
 Risk Management in Sterile Facilities
Jos C. Menezes coordinates a Systems
Engineering group with eight PhD students
and two post-Doctorates at the Technical
University of Lisbon, Centre for Biological
and Chemical Engineering. He has been work-
ing in Process Analytical Technology, namely
monitoring, chemometrics and multivariate
data analysis for more than 10 years. He has
a chemical engineering degree, a Masters in catalysis, and
a PhD in biochemical engineering and one post- Doctorate
in advanced monitoring of bioprocesses. He has been a Pro-
fessor at the Technical University of Lisbon for the past 10
years and has recently started his own company to market
PAT implementations in the industry. He can be reached by
email: cardoso.menezes@ist.utl.pt.
Institute for Biotechnology and Bioengineering, Centre for
Biological and Chemical Engineering, IST, Technical University
of Lisbon, Av. Rovisco Pais, 1049-001 Lisboa, Portugal.
Copyright ISPE 2010

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